SUBMISSION GUIDELINES

• Template and primer are submitted separately. 
• Template:
  • We require a minimum of 10 µL of DNA sample at the concentration specified in the table below. 
  • DNA must be purified and quantified prior to submission. It is critical that the concentrations are measured carefully, as incorrect quantitation (whether higher or lower) will result in poor sequencing results. Please note that the required concentrations listed are based on UV absorbance / nanodrop and not fluorometric quantification. See also “How should I quantify my DNA prior to submission?” in our FAQ section: https://sequencing.ubc.ca/our-services-equipment/sanger-sequencing/sanger-faq
  • If multiple reactions use the same template, please submit these as separate reactions (i.e. one 10 µl aliquot per reaction).
  • For orders with fewer than 48 samples, submit your samples in 8-strip PCR tubes. ONLY label the side of the tubes with your initials and sample number. For orders with 48 or more samples, submit your samples in a 96-well PCR plate, filling in columns (e.g. A1 to H1). A maximum of 92 samples can be submitted in a 96-well plate to leave room for control samples.  We can provide you with strips or plates if needed.
• Primer:
  • If submitting custom primers, a minimum of 5 µL of primer at 5 pmol/µL is required per reaction.
  • Primers should be submitted in 1.5 mL eppendorfs, with the primer name labelled on the lid with indelible ink.
• Templates and primers should be resuspended in water or Tris (not TE), as EDTA will interfere with the ion concentration in the sequencing reaction.
• Recommended concentrations are listed in the table below.

*only required if a standard sequencing primer is not specified

Please read important information on template/primer considerations in our submission guidelines.

• Template and primer are submitted pre-mixed with water. Our staff add sequencing chemistry, perform cycling, post-reaction clean-up, and electrophoreses.
• We require 14 µL of sample, containing template and primer at the amounts specified in the table below, mixed with water as needed.
• It is critical that the concentrations are measured carefully, as incorrect quantitation (whether higher or lower) will result in poor sequencing results. Please note that the required concentrations listed are based on UV absorbance / nanodrop and not fluorometric quantification. See also “How should I quantify my DNA prior to submission?” in our FAQ section: https://sequencing.ubc.ca/our-services-equipment/sanger-sequencing/sanger-faq

• For orders with fewer than 48 samples, submit your samples in 8-strip PCR tubes. ONLY label the side of the tubes with your initials and sample number. For orders with 48 or more samples, submit your samples in a 96-well PCR plate, filling in columns (e.g. A1 to H1). We can provide you with strips or plates if needed. A maximum of 92 samples can be submitted in a 96-well plate to leave room for control samples
• Templates and primers should be resuspended in distilled water or 10 mM Tris pH 7-8, with no EDTA, as EDTA will interfere with the ion concentration in the sequencing reaction.

• User performs the cycle sequencing with ABI Big Dye 3.1 Cycle Sequencing kit, and submits cycled product for post-reaction cleaning, injection, and electrophoretic separation.
• We require 12 µL of cycled material.
• For orders with fewer than 48 samples, submit your samples in 8-strip PCR tubes. Label the side of the tubes with your initials and sample number. For orders with 48 or more samples, submit your samples in a 96-well PCR plate, filling in columns (e.g. A1 to H1). We can provide you with strips or plates if needed. A maximum of 92 samples can be submitted in a 96-well plate to leave room for control samples
• Templates and primers should be resuspended in distilled water or 10 mM Tris pH 7-8, with no EDTA, as EDTA will interfere with the ion concentration in the sequencing reaction.

• User performs the cycle sequencing and post-reaction clean-up, and submits samples for injection and electrophoretic separation.
• We require 12 µL of sample resuspended in water, not formamide.
• For orders with fewer than 48 samples, submit your samples in 8-strip PCR tubes. Label the side of the tubes with your initials and sample number. For orders with 48 or more samples, submit your samples in a 96-well PCR plate, filling in columns (e.g. A1 to H1). We can provide you with strips or plates if needed.
• Templates and primers should be resuspended in distilled water or 10 mM Tris pH 7-8, with no EDTA, as EDTA will interfere with the ion concentration in the sequencing reaction.

Reaction Set-up

BigDye® dilution mix can be purchased from SBC (please see services and pricing)

Cycling Protocol

(ramp rate = 1°C/sec)

hot start	96°C	1 min
25x	96°C	10 sec
50°C	5 sec
60°C	4 min
soak	4°C	8

 
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Reaction Set-up

Cosmid/BAC

BigDye® dilution mix can be purchased from SBC (please see services and pricing)

Cycling Protocol

hot start	96°C	1 min
50x	96°C	10 sec
50°C	5 sec
60°C	4 min
soak	4°C	8